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Notes: The human sepiapterin reductase (SPR) gene has been mapped at the PARK3 locus, which is related to the onset of Parkinson disease. The silkworm Bombyx mori body color mutant lemon (lem) has been associated with a lack of SPR activity; lem lethal is a homozygous lethal allele of lem. Genetic linkage analysis was performed with normal silkworm strain p50T, lem strain l70, and leml strain a65 to more closely examine the relationship with SPR. DNA from the F1 and F2 crosses were isolated using the Wizard® SV 96 Genomic DNA Purification System and the genome sequenced. (4017)

Notes: To study how mutations in the quinolone resistance determining region (QRDR) of Staphylococcus epidermidis may have a role in fluoroquinolone resistance, 138 samples of S. epidermidis were swabbed from the conjunctival sacs of 129 patients. These samples were cultured overnight in tryptic soy broth, and genomic DNA isolated using the Wizard® SV 96 Genomic DNA Purification System. One microliter of the isolated DNA was used in PCR for the QRDR genes (gyrA, gyrB, parC and parE). (3939)

Notes: The authors developed an assay to evaluate antiviral compounds for Epstein-Barr virus (EBV) infections. EBV DNA was isolated using the Wizard® SV 96 Genomic DNA Purification System, and 5µl was used for real-time PCR to quantify the viral DNA. Cytotoxicity of the antiviral compounds was assessed using treated, uninfected Akata cells compared to those with no treatment or infection. After three days when the viral DNA was harvested, cell viability was measured using the CellTiter-Glo® Luminescent Cell Viability Assay. (3761)

Notes: To genotype PTPH1 knock-out (KO), heterozygous (HET), and wild type (WT) mice, tail snips were digested overnight with proteinase K and the DNA trapped using the Wizard® SV 96 Genomic DNA Purification System. Genomic DNA was washed using the Wizard® SV Wash Solution and eluted in 200μl of water at 65°C. The protease was inactivated at 95°C and 2μl of DNA was used for PCR. (3739)

Notes: In this study, a real-time PCR assay coupled with a DNA extraction method were used to detect staphylococcal enterotoxin (SE)-encoding genes in milk, a source of staphylococcal food poisoning. Pasteurized milk prepared with known concentrations of Staphylococcus aureus was used to generate a standard curve; experimental samples were from a staphylococcal food-poisoning outbreak that occurred in Japan in June 2000. PCR inhibition was overcome by using the following DNA purification method: a sample of milk (100µl) was added to an equal volume of 0.2M sodium hydroxide and incubated at 37°C for 20 minutes. The alkaline-treated sample was neutralized using 10µl of 3M sodium acetate (pH 5.4), extracted with 1ml of petroleum ether, and centrifuged at 13,000 × g for 10 minutes at 25°C. The aqueous phase was transferred to a fresh tube and bacterial DNA was purified from the aqueous solution using the Wizard® SV Genomic DNA Purification System. (3677)

Notes: To examine genetic variation in Anopheles funestus, genomic DNA was extracted from single mosquito carcasses (minus the abdomen) using the Wizard® SV 96 Genomic DNA Purification System on a Beckman Coulter Biomek® FX workstation. The purified genomic DNA was diluted 1:10 in water to ~5ng/µl for PCR analysis. (3415)

Notes: To confirm that the rat IL-10 transgene under the control of a liver-specific promoter was expressed in mice, mouse tails were subjected to Proteinase K digestion and genomic DNA isolation using the Wizard® SV 96 Genomic DNA Purification System. PCR analysis was used to distinguish between mouse and rat IL-10. (3555)

Notes: To examine both the efficiency of gene targeting constructs and the ability to knockin or knockout a gene, recombinant associated adenovirus (rAAV) constructs for p53 and CTNNB1 were created. Either HCT116 or DLD1 human colorectal cancer cell lines were infected with the rAAV and selected for resistance to Geneticin®. After the drug-resistant colonies were grown for 3–4 weeks, the genomic DNA from each clone was isolated using the Wizard® SV 96 Genomic DNA Purification System and locus-specific integration assessed by PCR. (3556)

Notes: The authors of this paper describe testing and comparing five genomic DNA purification kits for their ability to isolate bacterial DNA from Neisseria meningitidis-, Streptococcus pneumoniae-, and Haemophilus influenzae-inoculated whole blood samples. The Wizard® SV 96 Genomic DNA Purification System was considered to be the best kit for its ease of use and ability to be automated on a Roboseq 4200 PE liquid-handling robot. The Wizard® SV 96 Genomic DNA Purification System produced good yields of high quality DNA that displayed sensitivity levels down to 2 genome copies when used as template in fluorescence-based PCR (3209)

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